in African Journal of Microbiology Research [=AJMR] (2014), 8(3), 297-303

Lactic acid bacteria termed probiotics have preventive as well as curative effects on several types of diarrhoea of different aetiologies. The main objective of this study was to screen lactobacilli ... [more ▼]

Lactic acid bacteria termed probiotics have preventive as well as curative effects on several types of diarrhoea of different aetiologies. The main objective of this study was to screen lactobacilli strains having probiotic traits, isolated from Algerian healthy children faeces on the purpose of using them further in children diarrheal illnesses. One hundred and twenty (120) lactic acid bacteria isolates were selected from faecal samples of healthy Algerian children aged between five and ten years. Gram positive rods and catalase negative bacteria (52 isolates) were screened, in vitro, for their probiotic potential properties including ability to survive in simulated gastro-intestinal conditions, adherence to Caco-2 cells and their antimicrobial activity. The results show that only five strains resisted in simulated gastric juice at pH 1.5 and pepsin. Four of them were resistant to simulated intestinal conditions at pH 8 and pancreatin and have a good adherence. In the end, three of them were retained as they display interesting probiotic profiles characterized by a strong antimicrobial effect against some intestinal pathogenic bacteria. They were identified by 16S rDNA sequencing as Lactobacillus plantarum F12, Lactobacillus brevis G6 and Lactobacillus paracasei B13. [less ▲]

The aim of this work was the isolation and cultivation of amylolytic protists living in the digestive tract of the termite Reticulitermes santonensis (Feytaud). A chrysophyte identified as ... [more ▼]

The aim of this work was the isolation and cultivation of amylolytic protists living in the digestive tract of the termite Reticulitermes santonensis (Feytaud). A chrysophyte identified as Poterioochromonas sp. was isolated in a special medium containing rice grains as a source of carbon and nitrogen. Then, the protist was grown in a medium containing starch as a carbon source, tryptone, and a phosphate buffer at different pH values (5, 6 and 7). Yeast extract was added or not. Ciprofloxacin was used to avoid the bacterial development. Other antibiotics were also tested but showed an inhibitive effect on the growth of Poterioochromonas sp. Yeast extract allowed reaching 1.9 (pH 5), 2.3 (pH 6) and 2.2 (pH 7) times higher final cell concentrations, and 2.8 (pH 5), 2.8 (pH 6) and 2.2 (pH 7) times higher biomass yields. The starch concentration did not decrease in the medium until 3 and 4 days of culture, with and without yeast extract, respectively. Eight days of culture were necessary for hydrolyzing the starch completely, with and without yeast extract. Maltose and maltotriose were detected in the culture media and were hydrolyzed progressively. Maximal maltose concentrations were 0.68, 0.66 and 0.51 g.l-1 in the medium containing yeast extract. Maltotriose concentrations were only 0.17, 0.14 and 0.12 g.l-1. Other glucose oligomers were also detected but in lower quantities. It was determined that the protist developed a weak amylase activity, particularly at a weakly acidic pH (5-6). Such a pH also allowed a better growth of the protist. A maximal amylase activity of 112 nkat.l-1 was measured with yeast extract at pH 5. No other enzymatic activity (protease, cellulase or xylanase) was detected except amylase. The degradation products of starch which were obtained by enzymatic hydrolysis allow the identification of α-amylase, amyloglucosidase and possibly β-amylase activities. [less ▲]

The rhizobacterium Pseudomonas putida BTP1 stimulates induced systemic resistance (ISR) in tomato. A previous work showed that the resistance is associated in leaves with the induction of the first enzyme ... [more ▼]

The rhizobacterium Pseudomonas putida BTP1 stimulates induced systemic resistance (ISR) in tomato. A previous work showed that the resistance is associated in leaves with the induction of the first enzyme of the oxylipin pathway, the lipoxygenase (LOX), leading to a faster accumulation of its product, the free 13-hydroperoxy octadecatrienoic acid (13-HPOT), 2 days after Botrytis cinerea inoculation. In the present study, we further investigated the stimulation of the oxylipin pathway: metabolites and enzymes of the pathway were analyzed to understand the fate of the 13-HPOT in ISR. Actually the stimulation began upstream the LOX: free linolenic acid accumulated faster in P. putida BTP1-treated plants than in control. Downstream, the LOX products 13-fatty acid hydroperoxides esterified to galactolipids and phospholipids were more abundant in bacterized plants than in control before infection. These metabolites could constitute a pool that will be used after pathogen attack to produce free fungitoxic metabolites through the action of phospholipase A2, which is enhanced in bacterized plants upon infection. Enzymatic branches which can use as substrate the fatty acid hydroperoxides were differentially regulated in bacterized plants in comparison to control plants, so as to lead to the accumulation of the most fungitoxic compounds against B. cinerea. Our study, which is the first to demonstrate the accumulation of an esterified defense metabolite during rhizobacteria-mediated induced systemic resistance, showed that the oxylipin pathway is differentially regulated. It suggests that this allows the plant to prepare to a future infection, and to respond faster and in a more effective way to B. cinerea invasion. [less ▲]

in The Plant journal : for cell and molecular biology (2014), 77(3), 404-417

Isocitrate lyase is a key enzyme of the glyoxylate cycle. This cycle plays an essential role in cell growth on acetate, and is important for gluconeogenesis as it bypasses the two oxidative steps of the ... [more ▼]

Isocitrate lyase is a key enzyme of the glyoxylate cycle. This cycle plays an essential role in cell growth on acetate, and is important for gluconeogenesis as it bypasses the two oxidative steps of the tricarboxylic acid (TCA) cycle in which CO2 is evolved. In this paper, a null icl mutant of the green microalga Chlamydomonas reinhardtii is described. Our data show that isocitrate lyase is required for growth in darkness on acetate (heterotrophic conditions), as well as for efficient growth in the light when acetate is supplied (mixotrophic conditions). Under these latter conditions, reduced acetate assimilation and concomitant reduced respiration occur, and biomass composition analysis reveals an increase in total fatty acid content, including neutral lipids and free fatty acids. Quantitative proteomic analysis by 14 N/15 N labelling was performed, and more than 1600 proteins were identified. These analyses reveal a strong decrease in the amounts of enzymes of the glyoxylate cycle and gluconeogenesis in parallel with a shift of the TCA cycle towards amino acid synthesis, accompanied by an increase in free amino acids. The decrease of the glyoxylate cycle and gluconeogenesis, as well as the decrease in enzymes involved in beta-oxidation of fatty acids in the icl mutant are probably major factors that contribute to remodelling of lipids in the icl mutant. These modifications are probably responsible for the elevation of the response to oxidative stress, with significantly augmented levels and activities of superoxide dismutase and ascorbate peroxidase, and increased resistance to paraquat. [less ▲]

The chemical cues released by many insect species, including agricultural pests, are used by predators and parasitoids to locate their prey or host. For instance, aphids excrete honeydew, which contains ... [more ▼]

The chemical cues released by many insect species, including agricultural pests, are used by predators and parasitoids to locate their prey or host. For instance, aphids excrete honeydew, which contains bacteria that produce semiochemicals. Ladybeetles and hoverflies use these semiochemicals to locate the colonies of prey aphid species. One bacterium (Staphylococcus sciuri) has been identified in the honeydew of the pea aphid, Acyrthosiphon pisum. This bacterium is responsible for the production of kairomonal substances, which guide hoverflies to aphid colonies. In the present study, we cultivated S. sciuri, and used solid-phase microextraction (SPME) to confirm the ability of this bacterium to produce 3-methyl-2-butenal and 3-methyl-2-butenoic acid, which previous studies have demonstrated as being the two semiochemicals that exhibit kairomonal activity. We subsequently conducted field experiments to evaluate the efficiency of two solutions as biological products to control aphid populations inhabiting potato plants; the first solution contained a suspension of living S. sciuri, and the second solution contained a mixture of the two semiochemicals produced by this bacterium. While the semiochemical solution did not lead to a significant reduction in aphid number, potato plants treated with the S. sciuri solution were infested with 28% less aphids compared to untreated plants. This study demonstrates the potential of using naturally occurring bacteria as a form of biological control of aphid infestations in agricultural management. [less ▲]

The gammaproteobacterium Serratia symbiotica is one of the major secondary symbionts found in aphids. Here, we report the draft genome sequence of S. symbiotica strain CWBI-2.3T, previously isolated from ... [more ▼]

The gammaproteobacterium Serratia symbiotica is one of the major secondary symbionts found in aphids. Here, we report the draft genome sequence of S. symbiotica strain CWBI-2.3T, previously isolated from the black bean aphid Aphis fabae. The 3.58-Mb genome sequence might provide new insights to understand the evolution of insect-microbe symbiosis. [less ▲]

BACKGROUND: Whereas multi-species biofilm reactors are commonly used for the treatment of liquid and solid wastes, new strategies are progressing for the development of single species biofilm for the ... [more ▼]

BACKGROUND: Whereas multi-species biofilm reactors are commonly used for the treatment of liquid and solid wastes, new strategies are progressing for the development of single species biofilm for the production of high-value metabolites. Technically, this new concept relies on the design of bioreactors able to promote biofilm formation and on the identification of the key physico-chemical parameters involved in biofilm formation. RESULTS: An experimental setting comprising a liquid continuously recirculated on a metal structured packing has been used to promote Bacillus subtilis GA1 biofilm formation. The colonization of the packing has been visualized non-invasively by X-ray tomography. This analysis revealed an uneven, conical, distribution of the biofilm inside the packing. Compared with a submerged culture carried out in a stirred tank reactor, significant modification of the lipopeptide profile has been observed in the biofilm reactorwith the disappearance of fengycin and iturin fractions and an increase of the surfactin fraction. In addition, considering the biofilm reactor design, no foam formation has been observed during the culture. CONCLUSIONS: The configuration of this biofilm reactor set-up allows for a higher surfactin production by comparison with a submerged culture while avoiding foam formation. Additionally, scale-up could easily be performed by increasing the number of packing elements. [less ▲]

This report is to our knowledge the first to study plant growth promotion and biocontrol characteristics of Bacillus isolates from extreme environments of Eastern Algeria. Seven isolates of 14 (50 %) were ... [more ▼]

This report is to our knowledge the first to study plant growth promotion and biocontrol characteristics of Bacillus isolates from extreme environments of Eastern Algeria. Seven isolates of 14 (50 %) were screened for their ability to inhibit growth of some phytopathogenic fungi on PDA and some roots exudates. The bacteria identification based on 16S r-RNA and gyrase-A gene sequence analysis showed that 71 % of the screened isolates belonged to Bacillus amyloliquefaciens and the rest were closely related to B. atrophaeus and B. mojavensis. Most of them had high spore yields (22 x 108-27 x 108 spores/ml). They produced protease and cellulase cell wall-degrading enzymes while the chitinase activity was only observed in the B. atrophaeus (6SEL). A wide variety of lipopeptides homologous was detected by liquid chromatography-electrospray ionization-mass spectrometry analysis. Interestingly, some additional peaks with new masses were characterized, which may correspond to new fengycin classes. The isolates produced siderophores and indole-3- acetic acid phytohormone. The greenhouse experiment using a naturally infested soil with Sclerotonia sclerotiorum showed that the B. atrophaeus (6SEL) significantly increased the size of the chickpea plants and reduced the stem rot disease (P < 0.05). These results suggest that these isolates may be used further as bio-inoculants to improve crop systems. [less ▲]

A horizontal tubular fixed bed bioreactor (HFBR) and an anaerobic biodisc-like reactor (AnBDR) were designed to both fix Clostridium biomass and enable rapid transfer of the hydrogen produced to gas phase ... [more ▼]

A horizontal tubular fixed bed bioreactor (HFBR) and an anaerobic biodisc-like reactor (AnBDR) were designed to both fix Clostridium biomass and enable rapid transfer of the hydrogen produced to gas phase in order to decrease the strong effect of H2 partial pressure and H2 supersaturation on the performances of Clostridium strains. The highest H2 production rate (703 mL H2/L.h) and yield (302 mL/g glucose consumed i.e. 2.4 mol/mol) with the pure culture were recorded in the AnBDR with 300 mL culture medium (total volume 2.3 L) at pH 5.2 and a glucose loading rate of 2.87 g/L.h. These results are about 2.3 and 1.3-fold higher than those achieved in the same bioreactor with 500 mL liquid medium and with the same glucose consumption rate. Therefore, our experimentations and a short review of the literature reported in this paper emphasize the relevance of performing bioreactors with high L/G transfer. [less ▲]

Increased demand and progressive depletion of fossil fuels, and worldwide concerns about greenhouse gas emissions have resulted in the development of promising technologies for renewable energy production ... [more ▼]

Increased demand and progressive depletion of fossil fuels, and worldwide concerns about greenhouse gas emissions have resulted in the development of promising technologies for renewable energy production. Therefore, potential alternatives for energy generation are intensively studied. One option is the use of biomass feedstock for the production of biogas through anaerobic digestion. This process is a biochemical technological process for the treatment of a wide range of feedstocks (e.g. organic fraction of municipal waste, animal manure and slurry, agricultural crops, etc.) to produce methane-rich biogas which can be used in replacement of fossil fuels in both heat and power generation and as a vehicle fuel. However, there are critical issues, which need to be addressed to make the production of bio-methane techno-economically viable and ecologically acceptable. One of the most important issues is the effect of trace metals addition on anaerobic digestion. These metals can be stimulatory, inhibitory, or even toxic for biochemical reactions, depending on their concentrations. As reported in literature, Ni, Co and Fe are all involved in the methane production biochemical process and serve as cofactors in enzymes which are involved in the biochemistry of methane formation (Zandvoort et al., 2006). Recently, enormous interest has been focused on biological applications of metal nanoparticles NPs due to their small size, high specified surface and their great potential in application to many science fields. The most studied process concerns zero valent palladium and iron NPs improving anaerobic biodegradation of chlorinated hydrocarbons (Windt et al., 2005). Moreover, investigation carried out in our lab showed that iron NPs encapsulated in silicate matrix may enhance hydrogen production by Clostridium butyricum (Beckers et al., 2013). Nevertheless the influences of metal NPs on methane producing anaerobic digestion have seldom been investigated. The present work investigates the enhancement effect of seven different metal NPs on methane production during the thermophilic anaerobic digestion. NPs of Cu, Pd, Pt, Ni, Co, Ag and Fe encapsulated in porous silica (SiO2) to prevent their coagulation and agglomeration, were added at concentration of 10-5mol/L in batch test (125ml serum bottles containing 70mL culture medium with 5g/L acetate monohydrate as the sole carbon substrate. Nickel, cobalt and iron NPs improved methane production from acetate. To confirm the previous results, the NPs were tested at different concentrations (10-4, 10-5, and 10-6 mol/L) with starch and glucose substrates. The results show that the impact increases with the increase of NPs concentrations up to 10-4 mol/L. The modified Gompertz equation was applied to describe the effect of NPs on anaerobic digestion. According to this model, the kinetic of methane production was particularly affected by nanoparticles addition. The values of the maximum methane production rate MPR (ml/day) was significantly higher 72.5% with nickel NPs at a concentration of 10-4 mol/L than the control without NPs. [less ▲]

Generally, an antibiotic is thought to have a role in antagonism simply because the producing strain is known to exhibit a potential for pathogen growth inhibition. Some genetic approaches such as PCR ... [more ▼]

Generally, an antibiotic is thought to have a role in antagonism simply because the producing strain is known to exhibit a potential for pathogen growth inhibition. Some genetic approaches such as PCR using specific primers or genome mining using known sequence data of close relatives are also used. Nevertheless, none of these methods allows stating for a link between a specific compound and the observed antagonism. Yet MALDI Mass Spectrometry Imaging (MSI) is a powerful tool to decipher the chemical messengers exchanged by two protagonists [1,2,3;]. Tandem mass spectrometry (MS/MS) may be also used, either on extracts [2,3] or directly on the microbial colonies [4]. The presentation will thus be focused on two examples of application of MALDI MSI combined to in situ tandem mass spectrometry. The first presented case will be the antagonism between soilborne strain Paenibacillus polymyxa Pp56 and the fungal phytopathogen Fusarium oxysporum. Using MALDI MSI, we were able to precisely localize each detected antibiotic, allowing discriminating which LI-F lipopeptides (fusaricidin) were really active against the pathogen progression. Besides, the use of in situ MS/MS allowed us to sequence the peptide moiety of several LI-F lipopeptides, showing that some of them are actually a mixture of several forms. The second example concerns the metabolites that are released by Bacillus amyloliquefaciens S499 cells following their inoculation on 7 days old tomato roots. We developed specific bioassays for time-course monitoring by MALDI MSI. First analyses revealed an efficient secretion of surfactin by Bacillus cells after 3 days when colonization as biofilm-structured populations is well established. Even if the composition of antibiotic mixture does not greatly evolve over time, after long incubation periods (32 or 35 days post inoculation), new series of compounds are detected in the tomato root -surrounding medium. Structural analysis based on exact mass measurements and MS/MS experiments, performed directly on the semi-solid agar medium, allowed us to identify these compounds as new variants of surfactins. [1] Barger, S., et al., Anton Leeuw Int J G, 2012, 102, 435-445. [2] Hoefler, B. C., et al,. Natl Acad Sci USA, 2012, 109, 13082-13087. [3] Moree, W. J., et al., Natl Acad Sci USA, 2012, 109, 13811-13816. [4] Debois, D., et al., J Am Soc Mass Spectrom. 2013, 24, 1202-1213 [less ▲]

Morocco is continuously looking for developping Food industry through various programs (Moroccan Green Plan, National Initiative for Human Development, Emergence Plan. etc). It remains one of the most ... [more ▼]

Morocco is continuously looking for developping Food industry through various programs (Moroccan Green Plan, National Initiative for Human Development, Emergence Plan. etc). It remains one of the most major producers of fruit and vegetables countries (1.8 to 3 million tons, APEFEL 2012). The factual and numerical analysis of Moroccan fruit production showed that more than 30% of fruit production is lost during post harvest. The biochemical characterization of these local products (Dates, Cactus and Apples) reveals a rich composition of sugars and minerals needed for the growth of contamination microorganisms at the site of production. The objective of this study is to valorize post harvest fruit using biotechnological processes through the selection of endogenous and endemic microbial strains to prepare syrups, vinegars and starters for industrial use. Twenty-two samples (fruits, traditional alcohols and vinegars) from different regions of Morocco were used as substrates for isolation of bacteria and yeasts. Microbial isolates were there subjected to a biochemical, metabolic and molecular study for the identification and characterization. [less ▲]

The aim of this work was to isolate enzyme-producing microorganisms from the tract of the termite Reticulitermes santonensis. The microorganisms were extracted from the guts and anaerobic (CO2 or CO2/H2 ... [more ▼]

The aim of this work was to isolate enzyme-producing microorganisms from the tract of the termite Reticulitermes santonensis. The microorganisms were extracted from the guts and anaerobic (CO2 or CO2/H2) and micro-aerobic atmospheres were used to stimulate growth. Three different strategies were tried out. First, the sample was spread on Petri dishes containing solid media with carboxymethylcellulose, microcrystalline cellulose or cellobiose. This technique allowed us to isolate two bacteria: Streptomyces sp. strain ABGxAviA1 and Pseudomonas sp. strain ABGxCellA. The second strategy consisted in inoculating a specific liquid medium containing carboxymethylcellulose, microcrystalline cellulose, or cellobiose. The samples were then spread on Petri dishes with the same specific medium containing carboxymethylcellulose, microcrystalline cellulose, or cellobiose. This led to the isolation of the mold Aspergillus sp. strain ABGxAviA2. Finally, the third strategy consisted in heating the first culture and spreading samples on agar plates containing rich medium. This led to the isolation of the bacterium Bacills subtilis strain ABGx. All those steps were achieved in controlled atmospheres. The four enzyme-producing strains which were isolated were obtained by using a micro-aerobic atmosphere. Later, enzymatic assays were performed on the four strains. Streptomyces sp. strain ABGxAviA1 was found to produce only amylase, while Pseudomonas sp. strain ABGxCellA was found to produce β-glucosidase as well. Aspergillus sp. strain ABGxAviA2 showed β-glucosidase, amylase, cellulase, and xylanase activities. Finally, Bacillus subtilis strain ABGx produced xylanase and amylase. [less ▲]